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Formaldehyde-assisted Isolation of Regulatory Elements to Measure Chromatin Accessibility in Mammalian Cells
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JoVE Journal Genetica
Formaldehyde-assisted Isolation of Regulatory Elements to Measure Chromatin Accessibility in Mammalian Cells

Formaldehyde-assisted Isolation of Regulatory Elements to Measure Chromatin Accessibility in Mammalian Cells

DOI:
10.3791/57272-v

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08:08 min

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April 02, 2018

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, , , ,
1Department of Oncohematology and Genetics. Institute of Biomedicine of Seville (IBiS),University Hospital Virgen del Rocío, 2Institute of Biochemistry, Medical Faculty, Friedrichstrasse 24, Member of the German Center for Lung Research,Justus-Liebig-University

Capitoli

  • 00:04Titolo
  • 01:03Formaldehyde Crosslinking and Cell Harvesting
  • 02:18Cell Lysis and Chromatin Fragmentation
  • 03:25De-crosslinking of Control DNA
  • 04:04Phenol: Chloroform Purification of DNA
  • 06:44Results: Chromatin Accessibility Quantified
  • 07:29Conclusion

Summary

Traduzione automatica

Traduzione automatica

The plasticity of nuclear architecture is controlled by dynamic epigenetic mechanisms including non-coding RNAs, DNA methylation, nucleosome repositioning as well as histone composition and modification. Here we describe a Formaldehyde-Assisted Isolation of Regulatory Elements (FAIRE) protocol which allows the determination of chromatin accessibility in a reproducible, inexpensive, and straightforward manner.

Tags

Keywords: Chromatin AccessibilityFormaldehyde-assisted IsolationRegulatory ElementsMammalian CellsDNA-protein Cross-linkingChromatin RemodelingFAIRESonicationChromatin Fragmentation

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